Strategies to Improve Quantitative Proteomics: Implications of Dimethyl Labelling and Novel Peptide Detection
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In quantitative proteomics, many of the LC-MS based approaches employ stable isotopic labelling to provide relative quantitation of the proteome in different cell states. In a typical approach, peptides are first detected and identified by tandem MS scans prior to quantifying proteins. This provides the researcher with a large amount of data that are not useful for quantitation. It is desirable to improve the throughput of current approaches to make proteomics a more routine experiment with an enhanced capacity to detect differentially expressed proteins. This thesis reports the developments towards this goal, including an assessment of the viability of stable dimethyl labelling for comparative proteomic measurements and the evaluation of a dynamic algorithm called Parallel Isotopic Tag Screening (PITS) for the detection of isotopically labelled peptides for quantitative proteomics without the use of tandem MS scans.